Composition of halogenated hydroxy-diphenyl ethers

ABSTRACT

Halogen-2-hydroxy-diphenyl ethers are useful in bactericidal compositions and method in the protection of organic materials. Illustrative compounds are 2&#39;&#39;,4,4&#39;&#39;,5-tetrachloro-2hydroxydiphenyl ether, 4,4&#39;&#39;,5-trichloro-2-hydroxydiphenyl ether, 2&#39;&#39;,4,4&#39;&#39;-trichloro-5-bromo-2-hydroxydiphenyl ether and 4,4&#39;&#39;dichloro-5-bromo-2-hydroxydiphenyl ether.

United States Patent Model et al.

COMPOSITION OF HALOGENATED HYDROXY-DIPHENYL ETHERS Inventors: ErnstModel, Basel; Jakob Bindler,

Riehen, both of Switzerland Assignee: Ciba-Geigy Corporation, Ardsley,

Notice: The portion of the term of this patent subsequent to Dec. 21,1988, has been disclaimed.

Filed: Sept. 23, 1970 Appl. No.: 74,896 7 Related US. Application DataContinuation-impart of Ser. No. 627,603, Apr. 3, 1967, Pat. No.3,629,477, which is a continuation-in-part of Ser. No. 570,742, Aug. 8,1966,

Pat. No. 3,506,720, which is a continuation-in-part of Ser Ilo. 345,080,Feb. 17, 1964, aban do d.

US. Cl 424/304, 424/330, 424/340,

424/341, 252/106, 252/107 Int. Cl A01n 9/20 Field of Search 424/304,330, 340, 341

1*Mar. 26, 1974 [56] References Cited UNITED STATES PATENTS 2,950,3258/1960 Britton et a1 260/613 X 3,405,184 10/1968 Widiger, Jr. et al.260/613 3,417,146 12/1968 Linn et al. 260/613 X OTHER PUBLICATIONSChemical Abstracts Vol. 47 (1953) 7028.

Primary ExaminerAlbert T. Meyers Assistant ExaminerFrederick E. WaddellAttorney, Agent, or Firm-Karl F. Jorda; Martin J. Spellman [57] ABSTRACT7 Claims, No Drawings COMPOSITION OF HALOGENATED HYDROXY-DIPHENYL ETHERSplication Ser. No. 627,603, now U.S. Pat. No.'

3,629,477, filed Apr. 3, 1967, which is a continuationin-part of ourapplication Ser. No. 570,742, filed Aug.

- 8, 1966, now issued as Pat. No. 3,506,720, which is acontinuation-in-part of our application Ser. No.

. 345,080, filed Feb. 17, 1964, now abandoned.

DESCRIPTION OF THE INVENTION The present invention relates to the use ofnovel halogenated hydroxy-diphenyl ethers for the control ofmicroorganisms, and for the protection of organic materials and articlesfrom microorganisms particularly from bacteria and especially frominfestation with bacteria and growth of bacteria thereon, moreespecially, in a first aspect, for the disinfection of personal andhousehold linen, and for the protection of such materials from growth ofmicroorganisms thereon, and as bacteriostatic agents in bactericidalcompositions, for instance, washing agents and liquors.

Such control of microorganisms and, particularly bacteria, and moreespecially disinfection of a substrate normally permitting undesirablegrowth of bacteria thereon and/or protection of said substrate againstsuch growth, consists essentially of applying to said substrate adisinfecting and bacterial growth-inhibiting amount of a compound of theformula I:

wherein X is chlorine, bromine or hydrogen X is chlorine or bromine X ishydrogen, chlorine or bromine X is chlorine, bromine, alkyl having 1 to3 carbon X is hydrogen, chlorine, bromine, methyl, trichloron is l or 2.

Among these compounds of the formula I are two groups. The first ofthese groups has the formula I with the definition for X, as chlorineor/and bromine, X as chlorine or bromine, X as hydrogen, chlorine orbromine, methyl and trifluoromethyl'and n as l.

The preferred compounds of this group are those wherein X is in theposition 5 of the nucleus B. ln the most preferred compounds are Xchlorine or bromine and X hydrogen, chlorine or bromine. The second ofthese groups has the formula I with the definition of X as hydrogen, Xchlorine or bromine, X hydrogen, chlorine or bromine, X, as chlorine,bromine, methyl or NH and X,, as hydrogen, chlorine, bromine, methyl,trifluoromethyl or NHg and n as l or 2. Among the second group are twosubgroups. In the first of these subgroups n is defined as 1 and X isfixed to position 5 of the nucleus B. In the most preferred compounds ofthe first sub-group is X, chlorine and bromine and X is hydrogen,chlorine and bromine. In the second of the mentioned sub-groups n isdefined as 2, whereby those compounds are preferred having X fixed tothe positions 3 and 5 of the nucleus B. Most preferred are in thissub-group the compounds with X, as chlorine and bromine and X ashydrogen, chlorine and bromine.

Preferred used compounds of the present invention are 2 ,4, 4'5-tetrachloro-Z-hydroxydiphenylether, 4,4,5-trichloro-2-hydroxydiphenylether, 2' ,4, 4';trichloro-S-bromo-2-hydroxydiphenylether and 4,4-dichloro-5-bromo-2-hydroxydiphenylether.

Compositions according to the invention which contain a compound fallingunder Formulas I as active ingredient, in a bacteria growth-inhibitingamount, are distinguished by slight toxicity for warm blooded animalsand, in conventionally used concentrations, do not irritate the skin.They are bactericidally effective both against gram positive as well asgram negative bacteria, for example, against Bacillus mesientericus,Sarcina spec. and particularly against forms of Coli suchas againstEscherichia coli 96 and other gram negative organisms. A furtheradvantage of the halogen-ooxydiphenyl ethers used according to theinvention is their colorlessness or slight inherent color. This propertyenables them to be used for many purposes for which it is not possibleto use strongly solored known bactericidal compounds.

The above described compounds are not soluble in water but they aresoluble in dilute sodium and potassium hydroxide solutions and/or in allpractical organic solvents. Because of this solubility, they can be usedunder a first aspect according to the invention in very many ways forthe combatting of microorganisms, particularly of bacteria, and for theprotection of organic materials and objects from attack bymicroorganisms.

They can thus be incorporated directly into the material to beprotected, for example in material having a synthetic resin basis, aspolyamides and polyvinylchloride in paper treatment liquors, in printingthickeners made from starch or cellulose derivatives, in lacquers andpaints which contain, e.g. casein, in cellulose, in viscose spinningmass, in paper, in animal mucilages or oils, in permanent dressingshaving a basis of polyvinyl alcohols, in cosmetic articles such as insoaps, e.g. in hand or toilet soap, in ointments or powders. They canalso be added to preparations of inorganic or organic pigments for thepainting industry, plasticisers, etc.

Moreover, the above-described compounds of Formulas I can be used in theform of their organic solutions, e.g. as so-called spray or as drycleaners. As organic solvents, preferably those not miscible with waterare used, in particular, petroleum fractions, but also water misciblesolvents can be used such as low alcohols, e.g. methanol or ethanol orethylene glycol monomethyl or monoethyl ether.

In addition, they can be used with wetting or dispersing agents in theform of their aqueous dispersions, e.g. for the protection of substanceswhich tend to rot, such as for the protection of leather, paper etc.

Solutions or dispersions of active ingredient which can be used for theprotection of these materials advantageously have a content of activeingredient of at least 0.001 g/liter.

A preferred use for the diphenyl ether derivatives consists indisinfecting goods which are washed, and protecting such goods fromattack by microorganisms. For this purpose, either washing or rinsingliquors are used which contain the diphenyl ethers advantageously inconcentrations of about 1 to 200 parts per million calculated on theliquor.

As wash-active substances, the washing liquors contain, for example,anion active compounds such as aromatic sulfonic acids substituted bylipophilic groups or their water soluble salts such as the sodium saltof dodecyl benezene sulfonic acid, or water soluble salts of sulfuricacid monoesters of highter molecular alcohols or their polyglycolethers, e.g. soluble salts of dodccyl alcohol sulfate, or of dodecylalcohol polyglycol ether sulfate, or alkali metal salts of higher fattyacids (soaps), also nonionogenic wash-active substances such aspolyglycol ethers of higher fatty alcohols, polyglycol ethers of highermolecular alkylated phenols as well as so-called amphoteric wash-activesubstances such as reaction products of the alkali metal salts of lowhalogen fatty acids and polyalkylene polyamines containing lipohilicradicals, e.g. lauryl diethylenetriamine. In addition the liquor canalso contain the usual auxiliaries such as water soluble perborates,polyphosphates, carbonates, silicates, optical brightening agents,plasticisers,'salts having an acid reaction such as ammonium or zincsilicofluoride or certain organic acids such as oxalic acid, alsodressings such as, e.g. those having a basis of synthetic resin orstarch.

Chiefly, organic fibers are meant by goods which can be disinfectedwiththe washing or rinsing liquors according to the invention,containing the abovedescribed active compounds, namely those of naturalorigin such as cellulosic fibers, e.g. cotton, or polypeptide fibers,e.g. wool or silk, as well as fibers of synthetic origin such aspolyamide, polyacrylonitrile or polyester fibers or mixtures of theaforesaid fibers.

In the concentrations mentioned above, the diphenyl ether derivativesusable according to the invention disinfect the wash liquor as well asthe goods to be washed therein substantially freefrom Staphilococcusaureus or Coli and other bacteria, and these substrates remain free fromthese bacteria for a long time even after exposure to light of theactive ingredient or of the goods treated therewith. They differ fromother bactericidally active compounds particularly in their stability tolight on the goods treated therewith.

In a second aspect the invention concerns the protection of cellulosicmaterials such as wood and plants from the attack of microorganisms,among them rotcausing fungi, phytopathogenic fungi and bacteria.

Such protection comprises the application to the surface of suchmaterials, or incorporation thereinto, of a compound of the formula I inan amount sufficient to inhibit the growth of such microorganisms asrotcausing and phytopathogenic fungi and bacteria.

According to a third aspect, this invention concerns more particularly'aprocess for combatting pathogenic bacteria in the intestinal system andthe urinal tract of warm-blooded animals, and generally, in allmammalia, consisting essentially of administering to a warmbloodedanimal suffering from an attack of pathogenic bacteria in one of thesaid organs a bacteria growthinhibiting amount of a compound fallingunder formula I preferably in combination with an inert carriervthereforof the type described in detail further below.

The compounds of formula I have an excellent growth-inhibiting action,for example, on the following gram positive and gram negative bacteria:Staphylococcus aureus Smith, Sraphylocuccus Iactis,

Escherichia coli, Bacillus pumilus, Bacillus subtilis, C0- rynebacteriumdiphtheriae, Clostridium botulinum, Clostridium buryricum, Clostridumwelchii, Clostridium tetani, Klebsiella pneumoniae, Alcaligenesfaecalis, Salmonella pullorum, Salmonella typhi, Salmonella paratyphi Aand B, Salmonelly typhi murium, Salmonella enteritidis, Shigelladysentariae, Shigellaflexneri, Brucella aborlus, Proteus mirabilis,Achromobacter spec, Serratia marcescens, Pasteurella pseudotuberculosis.

They also inhibit the growth of the following pathogenic fungi:

Trichophylon mentagrophytes, Trichophyton rubrum, Trichophyton tonsuransvar. sabourandi, Trichophylon schonleini, Trichophyton quickeanum,Microsporon canis, Microsporon gypseum, Blaslomyced dermatidis,Sporotrichum schenkii, Epidermophytonfloccosum, Alternaria tenuis,Botrytis cenerea.

The use of compounds of formula I as active ingredients, is particularlyvaluable for the healing of diseased conditions of the intestinal systemand urinal track of warm blooded animals particularly in combattingpathogenic fungi and bacteria, since they are eliminated from the bodyin substantially unchanged, active form and they possess a low toxity.

Suitable for the latter type of use are tablets for the disinfection ofthe mouth and throat as well as tablets and sugar coated tablets(dragees) for the disinfection of the intestinal system and urinaltract.

Moreover, compounds of formula I are also useful as active ingredientsfor disinfectants for the hands, for

Moreover, compounds of formula I are also useful as active ingredients,for disinfectants for the hands, for example in soaps, cosmetics,ointments for wounds, eye ointments, ointments for skin and other agentsfor external use.

The diphenyl ethers usable according to the invention are also veryactive against the bacterial flora causing perspiration odours and,therefore, because of their low toxicity, are suitable as deodorants forlinenand for incorporation into cleansing agents such as soaps orshampoos or as additives for cosmetics such as ointments or creams.

The diphenyl ethers usable according to the invention can also be usedin combination with other antimicrobially active substances, for examplethey can be used with halogenated salicylic acid alkyl amides andanilides, with halogenated diphenyl ureas, with halogenated benzoxazolesor benzoxazolones, with polychlorohydroxydiphenyl methanes, withhalogendihydroxydiphenyl sulphides, with bactericidal 2-imino-imidazolidines or -tetrahydropyrimidines or with biocidalquarternary compounds or with certain dithiocarbamic acid derivativessuch as tetramethyl thiuram disulphide.

In addition, with some of the combinations mentioned of diphenyl ethersused according to the invention and other antimicrobial substances,there is a broadening of the range of action and/or a synergisticeffect.

The compounds used in the present invention can be produced byhalogenation of the corresponding halogenated and/or otherwisesubstituted 2-hydroxydiphenylether containing in the 5-position ahydrogen which can be prepared according to the copending application,Ser. No. 570,742, filed Aug. 8, 1966, by known processes, e.g. withelementary chlorine or bromine or with halogenating agents like sulfurylchloride, in order to receive the desired S-halogenated2-hydroxydiphenylethers of formula 1. Suitable 2-hydroxydiphenyletherswhich can be halogenated in 5-position from the copending applicationmentioned above are, for example: 2',4,4'-trichloro2-hydroxydiphenylether, 4,4'- dichloro-2-hydroxydiphenylether,4,4-dibromo-2- hydroxydiphenylether,4'-methyl-4-chloro-2-hydroxydiphenylether, 2 ',4,4,5'-tetrachloro-2-hydroxydiphenylether, 4,4'-dichloro-3-trifluoromethyl-2-hydroxydiphenylether, 4,4-dichloro-3 -methyl-2- hydroxydiphenylether,4'-methoxy-4-chloro-2-hydroxydiphenylether,4,4'-dichloro-2'-cyano-2-hydroxydiphenylether,

In case of 2-hydroxydiphenylethers containing an amino group, the aminogroup is acetylated before the halogenatio n takes place and afterwardshydrolized to the corresponding halogenatedamino-2-hydroxydiphenylether. A suitable aminc-2-hydroxydiphenyletheraccording to the copending application, Ser. No.

of l-nitro-2-halogenobenzenes are vl-nitro-2,5- dichlorobenzene,l-nitro-2,3,S-trichlorobenzene, or l-nitro-2,S-dibromobenzene.

A third method according to the copending application mentioned beforeconsists in the condensation of l-alkoxy-Z-chlorobenzene or 1-alkoxy-2-bromobenzene with an alcali-metal-salt of a hydroxybenzene which may besubstituted by halogen, alkyl, or a trifluoromethyl group; thecondensation beingvperformed in the presence of copper (I) salts orcopper powder to form the corresponding o-alkoxydiphenylether andfinally the alkoxy group converted into the hydroxyl group. Halogenationwith chlorine, bromine or SO Cl of the2-hydroxydiphenylethers preparedin this process leads to the compounds of formula III.

The preparation of 2-hydroxy-diphenylethers of Formula lll containing anamino group according to the third method is modified in so far that theintermediate amino-2-alkoxy-diphenylether is acetylated and afterwardshalogenated and hydrolysed to the correspondingamino-2-alkoxy-3,5-dihalogenated hydroxydiphenylether or theamino-2-alkoxy-5-halogenated hydroxy-diphenylether, respectively, whichfinally, by opening of the methoxyether leads to the amino-2-hydroxy-diphenylether of Formula Ill. An example is:4,3,5-trichloro-2'-amino-2-hydroxydiphenylether. An other modificationconsists in the acetylation of an amino-2-hydroxy-diphenylether not yethalogenated in ring B. The halogenation leads to the acetylamino-3,5-dihalogeno-2-hydroxydiphenylether or theacetylamino-5-halogeno-2-hydroxy-diphenylether, respectively,

which then is hydrolised to the products of Formula llI.

The following non-limitative examples further illustrate the invention.

- EXAMPLE 1 in a three necked flash equipped with a stirrer, thermometerand a dropping funnel 25.5 g of 4,4'-dichloro- Z-hydroxy-diphenyletherare dissolved in 35 ml of glacial acetic acid. In the dropping funnel15.98 g of elementary bromine are dissolved in 20 ml of glacial aceticacid and added dropwise over one hour at 8 14 10C to the solution of4,4-dichloro-2-hyd'roxydiphenylether.

The reaction product was stirred for three hours at room temperature andthen added to 1.5 liter of icewater. The oily product which separates isstirred in the ice-water for 2 hours and solidifies slowly. Filtrationand drying in vacuum yields 32 g of a product melting at C. Afterrecrystallisation the 4,4-dichloro- 5-bromo-2-hydroxy-diphenylethermelts at 98 100C in a yield of 61.7 percent.

EXAMPLE 2 EXAMPLE 3 in a three necked flash equipped with stirrer,thermometer and dropping funnel 28.9 g of 2',4,4-trichloro-2-hydroxy-diphenylether are dissolved in 100 ml ofchlorobenzene. At a temperature of 25C 13.9 g of sulfuryl chloride areadded dropwise. Over night the temperature is kept at 90 l00C andfinally for three hours at reflux temperature. The reaction mixture iscooled down to60C and 'the solvent distilled off in vacuum. The residueis then recrystallized threetimes in petroleum ether yielding thedesired 2',4,4,5- tetrachloro-2-hydroxy-diphenylether with the m.p. of85 87C. 1 1

EXAMPLE 4 By substituting 2',4,4-trichloro-2-hydroxydiphenylether with4,4f-dichloro-Z-hydroxydiphenylether as described in Example 3andchlorination, the new 4,4,5-trichloro-2-hydroxy-diphenylether withthe m.p. of 85 87C was prepared.

Further 5-chloro-2-hydroxy-diphenylethers which can be prepared inanalogy to Example 3 are:

4,4-dibromo-5-chloro-Z-hydroxy-diphenylether, 4,5-dichloro-4'-methyl2-hydroxydiphenylether, 2,4,4',5,5'-penta-chloro-diphenylether,2-amino-4,4,5-trichloro-Z-hydroxy-diphenylether.

EXAMPLE a. 25 g of 4-chlorophenol are dissolved in 150 ml ofdimethylformamid. To this solution 13.2 g of powdered potassium-hydroxydand 0.2 of copper powder are added. The reaction mixture is heated to90C and at this temperature 37.4 g of l-methoxy-2-bromobenzene are addeddropwise within minutes. Heating is continued for hours at refluxtemperature. After cooling down to room temperature the reaction mixtureis poured in 2 liter of water containing 50 ml of 30 percentsodium-hydroxyd solution. The oily product is extracted with ether; theether phase washed neutral with water and the ether evaporated. Thecrude oil is distilled and the fraction with the b.p. of 107 1 10C at0.02 mm collected. Recrystallisation from petroleum ether yields4'chloro-2-methoxy-diphenylether with a m.p. of 50 52C.

b. 9.36 g of 4'-chloro-2-methoxy-diphenylether are dissolved in 40 ml ofglacial acetic acid. Then 25 ml of hydrobromic acid are added. Theresulting turbid 'reaction mixture is heated over night whereby a clearsolution is formed. After cooling down to room temperature the productof 4'- chloro-Z-hydroxy-diphenylether is precipitated with water,filtered off, dried and recristallized in petroleum ether to yield 6 g(68 percent of pure 4- chloro-2-hydroxy-diphenylether with the m.p. of82 84C.

c. 8.8 g of 4-chloro-2-hydroxy-diphenylether are dissolved in 70 ml ofglacial acetic acid. At a temperature of C 6.2 g of chlorine are bubbledinto the reaction mixture within about 90 minutes. This reaction isslightly exotherm. After stirring at room temperature over night thereaction product is poured into 400 ml of ice water. The crude productis extracted with benzene, washed neutral with water, the benzene phasedried over sodium sulfate, the benzene distilled off under reducedpressure and the yield of 12 g recrystallized in petroleum ether toreceive pure 3,4,5-trichloro-2-hydroxydiphenylether with the m.p. of 7981C.

Chlorination with only one equivalent of chlorine furnishes the4,S-dichloro-2-hydroxy-diphenylether.

With this same procedure the following compounds can be prepared:

4'-methyl-3,5-dichloro2-hydroxy-diphenylether,

4'-chloro-3-trifluorornethyl-3,5-dichloro-2-hydroxydiphenylether,3,4,3,5-tetrachloro-2-hydroxy-diphenylether.

Bromination according to Example 1 leads to the corresponding brominatedproducts:

4'-chloro-3,5-dibromo-Z-hydroxy-diphenylether,

(m.p. 96-98C) 4'-methyl-3,5-dibromo-2-hydroxy-diphenylether,

4-chloro-3-trifluoromethyl-3,5-dibromo-2-hydr0xydiphenylether,

4-chloro-5-bromo-2-hydroxy-diphenylether.

EXAMPLE 6 Agar Incorporation Test For each test the active substance tobe tested was added in graduated concentrations to nutrient agar (D-ifco) The test substance was first dissolved in Methyl Cellosolve(Methyl Cellosolve is a tradename of Union Carbide Corp.), and thefollowing concentrations of each of the test substances, calculated onthe weight of the nutrient agar, prepared:

0.1, 0.3, l, 3, 10, 30, and 300 parts per million (p.p.m.). Theresulting mixtures were then poured directly into sterile 100 mm Petridishes in amounts of about 20 ml per dish. After inoculation with thebacteria the agar plates were incubated for 48 hours at 37C. The lowestconcentration of the different test substances incorporated in thenutrient agar which inhibited all growth of the bacteria was thentabulated.

The following strains of bacteria were tested:

Staphylococcus aureus Staphaureus) Escherichia coli (E.c0li)Streptococcus faecalis Brevibacterium ammoniagenes Sarcina ureae Proteusvulgaris Bacillus subtilis Salmonella pullorum The results are compiledin the table below giving the minimal inhibitory concentration of testsubstances in the nutrient agar expressed in p.p.m., and compound A is4,4',5-trichloro-2-hydroxy-diphenylether, compound B is4,4-dichloro-5-bromo-2-hydroxy-diphenylether, compound C is2,4,4',5-tetrachloro-2-hydroxydiphenylether, and compound D is2,4,4-trichloro-5 bromo-2-hydroxy-diphenylether. A

Laundry Test Washed Bath Disinfection This test shows the disinfectanteffect of the test substances on infected laundry when the testsubstances are applied in the wash bath. There are three aspects to thistest: 7

l. the Germ Count of the wash bath is determined,

2. the textile samples are tested for the presence of any remainingbacteria by the Bacterial Growth Test, and

3. after being dried, the textile samples are tested for any residualsanitizing effect by the Inhibition Test.

This latter sanitizing effect is for example of special importance forhospital laundry, which is readily reinfected by germs in the air orfrom personnel handling the material before it is used again by thepatient. These tests were carried out on the gram negative bacterium E.coli and on the gram positive bacterium Staph. aureus.

1. Wash Raw, unfinished cotton fabric, which had not been treated withoptical brightening agents was inoculated with 24 hour-old strains ofthe test bacteria, which had been prepared on agar slants. 2 g of thisinoculated cotton fabric were added to each of the test wash bathscontaining 50 p.p.m. and 100 ppm. active ingredient respectively(calculated on the wash bath) and washed for minutes at 40C (liquorratio 1:20).

Biological Tests a. Germ Count of the Wash Bath A series of test agarplates were prepared to test for the presence of Escherichia colibacteria by adding 20 ml of nutrient agar according to McConkey, and totest for the presence of Staph.aureus by adding 20 ml of nutrient agarcontaining 0.5 percent by weight of potassium tellurite to 1 ml of thetest wash baths and pouring the mixture into Petri dishes. Afterincubation for 24 hours at 37C the colonies of germs were counted on theagar plates. The results are given in the table below:

Disinfection Effect in the WashBath germs/milliliter b. Disinfection ofthe Textile Material After removal of 1 ml sample of-the wash bath forthe germ count test, the cotton samples (2.5 cm in diameter) were eachrinsed twice with permutite water and placed on the agar plate preparedaccording to McCconkey, supra, and nutrient agar containing 0.5 percentby weight of potassium tellurite for tests with E.coli and Staph. aureusrespectively. The agar plates in Petri dishes were incubated for 24hours at 37C and the bacterial growth on the textile samples and on theagar plates was evaluated visually. The results are given in the tablebelow wherein represents growth under or/and on the textile sample nogrowth on and under the textile sample.

0. Inhibition Test (Residual sanitizing effect on the textile material).

Finally the remaining cotton samples were air-dried and tested for theirresidual disinfectant activity i'n-the inhibition test.

10 ml each of Bacto-Agar (Difco) were poured into Petri dishes. 10 ml ofmolten nutrient agar prepared as V stated under (a), and inoculated withE.coli and Staphaureus respectively were overlaid on each of theprepared Petri dishes containing the Bacto-Agar. The

dried cotton samples were laid on the plates which then were incubatedfor 24 hours at 37C. Thereafter the zones of growth inhibition weredetermined in millimeters. The results are compiled in the table below:

Residual Sanitizing Effect on the Textile Material I (zones ofinhibition in mm) Laundry Test Rinse Bath Disinfection This test issimilar to the wash bath disinfection test except that sterile textilematerial is used and the test bacteria as well as the test substanceswere both added to the third rinse bath and the following experimentsperformed: A

25 l. Germ Count Test of the rinse bath,

2, Bacterial Growth Test Disinfection of the textile material, 3.Residual Sanitizing Effect on the textile material in inhibition test. 7

1. Rinse The cotton samples were rinsed twice for three minutes each at20C with sterile permutite water (liquor ratio 1:20).

To the third rinse bath the active ingredient to be tested was added toprepare the concentrations of 25 ppm, 50 ppm and 100 ppm a.i. calculatedon the total rinse' bath. Then the bath was inoculated with E.coli andStaph. aureus and the cotton samples then rinsed a third time for threeminutes at 20.

2. Biological Tests a. Germ Count in the Rinse Bath 1 ml of the thirdrinse bath was added to 20 ml of nutrient agar prepared according toMcConkey, to test E.coli and to 20 ml of nutrient agar containing 0.5percent by weight of potassium tellurite to test Staph- .aureus. Theresulting mixtures were poured into Petri dishes and incubated for 24hours at 37C. The viabel germ count per milliliter of rinse bath wasdetermined 50 by counting the number of colonies formed on the agarplates. The results are compiled in the following table: 7

' Disinfection Effect in the Rinse Bath germs milliliter Compound E.coli 1 Staph. aureus 50 ppm 100 ppm 50 ppm 3 I00 ppm Control l0 l0 10 10b. Disinfection ,of the Textile Material After removal of 1 ml sample ofthe rinse bath for the 5 germ count test, the cotton samples (2.5 cm indiameter) were each placed on one agar plate prepared according toMcConkey, supra, thereby using nutrient agar and nutrient agarcontaining 0.5 percent by weight of potassium tellurite for the testswith Ecoli and 2. The process of claim 1, wherein in said compoundStaph.aureus, respectively. The agar plates were incubated for 24 hoursat 37C and then evaluated visually. X, and X are independently chlorineor bromine, The results are given in the table below, wherein X, ishydrogen, chlorine or bromine,

+ represents growth under or/and on the textile sam- X is chlorine,bromine, methyl, methoxy, cyano or ple amino, no growth on and under thetextile sample. X is hydrogen, chlorine, bromine, methyl, trifluoro-Disinfection Test of Textile Material Bacterial n g or ammo and GrowthTest 3. A process for combatting pathogenic bacteria in c d E s h theintestinal system and the urinal tract of warmompoun C0! lap aureus t 50ppm 100 ppm 50 ppm '00 ppm blooded animals, wh ch comprises orallyadmlmstermg a warm-blooded animal suffering from an attack of Ipathogenic bacteria in one of said organs a bacterially Cgrowth-inhibiting amount of a compound of the for- D mula Control g X5 I2)n c. lnhibmon Test X,O B -X (Residual sanitizing effect on the textilematerial). I Finally after the third rinse the remaining cotton sama 0Hples were air dried and tested for their residual disinfectant activityin the inhibition test. 10 ml each of Bacto I wherem Agar (Dlfco) werepoured mto Petri dishes. 10 ml of X and X are independently hydrogen,chlorine or molten nutrient agar prepared as stated under (a) andbromine inoculated with E.coli and Staph.aureus respectively wereoverlaid on each of the prepared agar plates. The dried cotton sampleswere put on the agar, which then were incubated for 24 hours at 37C.Thereafter the zones of growth inhibition were determined inmillimeters. The results are given in the following table:

X is chlorine or bromine X is chlorine, bromine, alkyl having 1 to 3carbon atoms, methoxy, cyano or amino X is hydrogen, chlorine, bromine,methyl, trifluoromethyl, methoxy, cyano or amino, and

n is l or 2. Residual Sanitizing Effect on the Cotton Samples 4. Theprocess of claim 3, wherein in said compound (Zones of inhibition in mm)X and X are independently chlorine or bromine, Compound E. coli Staph.aureus X is hydrogen, chlorine or bromine I 25 ppm 25 100 X is chlorine,bromine, methyl, methoxy, cyano or A 2 3 6 s amino B 2 3 3 8 40 X ishydrogen, chlorine, bromine, methyl, trifluorog g g I g I methylor'amino, and

n is 1.

Control 0 0 0 0 5. A process for combatting larvae of Attagenus andgrowth Anthrenus beetles which comprises applying to said +=gmmh larvaean inhibitory amount of a compound of the formula What we claim is: 1. Aprocess for combatting bacteria and phytopathogenic fungi whichcomprises applying to said bacteria X5 (Xm or fungi an inhibitory amountofa compound of the for- A mula X, on X5 011 X,- A -O B -X1 wherein X HX and X, are independently hydrogen, chlorine or bromine,

X is chlorine or bromine wherein X is chlorine, bromine, alkyl havingone to three car- X and X, are independently hydrogen, chlorine or batoms, h cyano amino bromine, X is hydrogen, chlorine,'bromine, methyl,trifluoro' X is chlorine or bromine, methyl, methoxy, cyano or amino,and X is chlorine, bromine, alkyl having one to three carn i l o 2,

bon atoms, methoxy, cyano or amino, 6. A composition for combattingbacteria, phyto- X is hydrogen, chlorine, bromine, methyl,trifluoropathogenic fungi and larvae of Attagenus and Anmethyl, methoxy,cyano or amino, and threnus beetles comprising an inert carrier and anin n is 1 or 2. hibitory amount of a compound of the formula 13 14 X (XmX is chlorine, bromine, alkyl having 1 to 3 carbon atoms, methoxy, cyanoor amino I 7 X is hydrogen, chlorine, bromine, methyl, trifluorot X; 7 V,7 (1H 7 H v '7 methyl, methoxy, cyano or amino, and

wherein 5 n is 1 or 2.

X1 and X3 are independently hydrogen chlorine or 7. The composition ofclaim 6 wherein said carrier is bromine, a pharmaceutically acceptableone.

X is chlorine or bromine

2. The process of claim 1, wherein in said compound X1 and X2 areindependently chlorine or bromine, X3 is hydrogen, chlorine or bromine,X4 is chlorine, bromine, methyl, methoxy, cyano or amino, X5 ishydrogen, chlorine, bromine, methyl, trifluoromethyl or amino, and nis
 1. 3. A process for combatting pathogenic bacteria in the intestinalsystem and the urinal tract of warm-blooded animals, which comprisesorally administering to a warm-blooded animal suffering from an attackof pathogenic bacteria in one of said organs a bacteriallygrowth-inhibiting amount of a compound of the formula
 4. The process ofclaim 3, wherein in said compound X1 and X2 are independently chlorineor bromine, X3 is hydrogen, chlorine or bromine X4 is chlorine, bromine,methyl, methoxy, cyano or amino X5 is hydrogen, chlorine, bromine,methyl, trifluoromethyl or amino, and n is
 1. 5. A process forcombatting larvae of Attagenus and Anthrenus beetles which comprisesapplying to said larvae an inhibitory amount of a compound of theformula
 6. A composition for combatting bacteria, phytopathogenic fungiand larvae of Attagenus and Anthrenus beetles comprising an inertcarrier and an inhibitory amount of a compound of the formula
 7. Thecomposition of claim 6 wherein said carrier is a pharmaceuticallyacceptable one.